OALib Journal期刊
Real-timePCR与常规PCR在Q热立克体检测中的比较
, PP. 1054-1056
Keywords: Q热立克次体 ,Q热 ,Real-time ,PCR ,常规PCR ,评价
Abstract:
目的比较与评价Real-timePCR与常规PCR,寻求适合于Q热患者早期诊断的方法。方法应用TaqMan探针定量PCR、SYBRGreen染料定量PCR、巢式PCR和普通PCR四种方法分别对T-A克隆的Q热立克次体基因htpAB片断进行灵敏性、特异性和重复性检测及评价。结果TaqManPCR方法的灵敏度分别为SYBRGreenPCR、巢式PCR和普通PCR的10倍、10倍和100倍。重复性测试TaqMan法Ct变异系数CV0.2%~3.0%,SYBRGreen法的CV0.3%~6.0％。TaqMan和SYBRGreenPCR均耗时约1h,巢式PCR耗时约2.5h,普通PCR耗时约1.5h。四种方法检测25种相关病原菌结果均为阴性。结论TaqMan探针定量PCR方法具有很好的特异性、灵敏性、重复性和可操作性,可用于Q热患者临床早期诊断。
References
[1] 俞东征主编.人兽共患传染病学[M]. 北京: 科技出版社,2009：657-668. [2] 张丽娟. 中国立克次体病监测及防治现状与展望 [J]. 疾病监测, 2007, 22(9): 577-579. [3] Zhang LJ, Fu XP, Zhang JS. Q fever endocarditis with multi-organ complication: a case report [J]. Chin Med J, 2006, 119 (18):1580- 1582. [4] Karakousis PC, TrucksisM, Dumler JS. Chronic Q fever in the United States [J]. J Clin Microbiol, 2006, 44(6): 2283-2287. [5] 魏文进,温博海. Q 热疫苗研究进展 [J]. 微生物学免疫学进展, 2004, 32(3): 64-68. [6] Fenollar F, Fournier PE, Raoult D. Molecular detection of Coxiella burnetii in the sera of patients with Q Fever endocarditis or vascular infection [J]. J Clin Microbiol, 2004, 42(11): 4919-4924. [7] 张晶波, 温博海, 陈梅玲, 等. 实时荧光定量PCR检测汉赛巴通体[J]. 中华流行病学杂志, 2007, 28(3): 277-281. [8] 亚红祥,赫兢,张丽娟,等. 贝氏柯克斯体实时荧光定量PCR方法的建立及对云南鼠标本检测 [J]. 传染病信息, 2009,22(6):345-347. [9] Kaltenboeck B, Wang C．Advances in real-time PCR: application to clinical laboratory diagnostics [J]．Adv Clin Chem, 2005(2),40: 219-259． [10] Kubista M, Andrade JM, Bengtsson M, et al. The real-time polymerase chain reaction [J]. MolAspects Med, 2006, 27(2-3): 95 -125. [11] Tse C, Capeau J. Real time PCR methodology for quantification of nucleic acids [J]. Ann Biol Clin,2003, 61(3):279-293. [12] Arya M, Shergill IS, Williamson M, et al. Basic principles of real-time quantitative PCR [J]. Expert Rev Mol Diagn,2005, 5(2):209-219. [13] Valasek MA, Repa JJ. The power of real-time PCR [J]. Adv Physiol Educ, 2005, 29(4): 151-159.
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